Objective: To construct the recombinant Ad. mda-7/IL-24 using Adeasy 1 system.
Methods: The mda-7/IL-24 DNA sequence was, by PCR, amplified from the plasmid pREP4-mda-7 and sub-cloned into the shuttle vector pAdTrack CMV. The resultant plasmid and pAdEasy 1 were used to co-transfer into the E. coli BJ5183 cells to undergo homologous recombination. The linearized recombinant plasmid DNA was transferred into 293 cells. Then the ovarian cancer drug resistant cell line OVCAR-8/TR was infected by the recombinant adenovirus, in which the expression of MDA-7/IL-24 protein was detected by Western-blot analysis.
Results: The recombinant Ad. mda-7/IL-24 was constructed successfully and approved by sequence analysis, electrophoresis and expression of MDA-7/IL-24 protein. All OVCAR-8/TR cells had been infected under concentration of Ad. mda-7/IL-24 was 10 microL. OVCAR-8/TR cell had the objective protein expression after infected.
Conclusion: The recombinant adenovirus Ad. mda-7/IL-24 is successfully constructed, which lays a foundation for studying mda-7/IL-24 gene in the field of ovarian cancer drug resistant.
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